Enzymes are highly specific catalysts used increasingly for applications that include fine-chemical synthesis, pharmaceuticals, food processing, detergent applications, biosensors, bioremediation, protein digestion in proteomic analysis, and biofuel cells. Despite the variety of enzymes and methods available, development of both stable and active enzyme systems remains a challenging issue in realizing successful use of enzymes for many practical applications. Recent attention has focused on use of nanostructured materials including mesoporous media, nanoparticles, carbon nanotubes, and nanofibers as enzymatic supports, as such materials provide large surface areas that can lead to high volumetric enzyme activity. Nanofibers offer a number of attractive features compared with other nanostructures. First, nanofibers do not have the same mass transfer limitations of other nanostructures such as mesoporous media due to their reduced thicknesses. Second, nanofibers are easily formed or processed into various structures such as non-woven mats, well-aligned arrays, and/or membranes—all with controllable compositions and sizes. However, loading capacity by known methods is limited to monolayers. Accordingly, new processes are needed that can further improve enzyme loading leading to increased overall enzymatic activity.